A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.

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A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.
Robertson, Adam B; Dahl, John A; Vågbø, Cathrine B; Tripathi, Pankaj; Krokan, Hans E; Klungland, Arne
Nucleic acids research 2011-02-07
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dc.contributor.authorRobertson, Adam Ben
dc.contributor.authorDahl, John Aen
dc.contributor.authorVågbø, Cathrine Ben
dc.contributor.authorTripathi, Pankajen
dc.contributor.authorKrokan, Hans Een
dc.contributor.authorKlungland, Arneen
dc.identifier.citationNucleic acids research 2011-02-07en
dc.description.abstractRecently, 5-hydroxymethylcytosine (5hmC) was identified in mammalian genomic DNA. The biological role of this modification remains unclear; however, identifying the genomic location of this modified base will assist in elucidating its function. We describe a method for the rapid and inexpensive identification of genomic regions containing 5hmC. This method involves the selective glucosylation of 5hmC residues by the β-glucosyltransferase from T4 bacteriophage creating β-glucosyl-5-hydroxymethylcytosine (β-glu-5hmC). The β-glu-5hmC modification provides a target that can be efficiently and selectively pulled down by J-binding protein 1 coupled to magnetic beads. DNA that is precipitated is suitable for analysis by quantitative PCR, microarray or sequencing. Furthermore, we demonstrate that the J-binding protein 1 pull down assay identifies 5hmC at the promoters of developmentally regulated genes in human embryonic stem cells. The method described here will allow for a greater understanding of the temporal and spatial effects that 5hmC may have on epigenetic regulation at the single gene level.en
dc.publisherOxford Journalsen
dc.subjectVDP::Medisinske Fag: 700::Klinisk medisinske fag: 750::Onkologi: 762en
dc.titleA novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.-
dc.typeJournal articleen
dc.typepeer revieweden
dc.contributor.departmentCentre for Molecular Biology and Neuroscience and Institute of Medical Microbiology, Oslo University Hospital, Rikshospitalet, Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, N-7489 Trondheim, Norway, Division of Molecular Biology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, The Netherlands and Institute of Basic Medical Sciences, University of Oslo, PO Box 1018 Blindern, N-0315 Oslo, Norway.en
dc.identifier.journalNucleic acids researchen
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